Methods for in vitro evaluation using functional engineered three-dimensional tissues with circumferential or helically oriented tissue structure

Abstract

Methods of evaluating functional activity of an engineered three-dimensional tissue, methods of identifying a compound that modulates tissue function, and methods for identifying a compounds that is useful for treating or preventing a disease affecting tissue function are provided herein. Some methods include providing or obtaining a three-dimensional tissue scaffold defining a lumen or a cavity, the tissue scaffold comprising one or more polymeric fibers each having a micron-scale or nanometer-scale diameter, at least some of the one or more polymeric fibers encircling the lumen or cavity at a helical angle with respect to a longitudinal axis of the lumen or cavity, at an azimuthal orientation with respect to the longitudinal axis of the lumen or cavity, or at both.

1 . A method comprising:
 providing or obtaining a three-dimensional tissue scaffold defining a lumen or a cavity, the tissue scaffold comprising one or more polymeric fibers each having a micron-scale or nanometer-scale diameter, at least some of the one or more polymeric fibers encircling the lumen or cavity at a helical angle with respect to a longitudinal axis of the lumen or cavity, at an azimuthal orientation with respect to the longitudinal axis of the lumen or cavity, or at both;   providing or obtaining cells growing on or in the three-dimensional tissue scaffold to form a three-dimensional tissue defining the lumen or the cavity;   affixing a first portion of the three-dimensional tissue to a support where the cells are grown on or in the three-dimensional tissue scaffold to form the three-dimensional tissue before the three-dimensional tissue is affixed to the support, or affixing a first end of the three-dimensional tissue scaffold to a support where the cells are grown on or in the three-dimensional tissue scaffold to form the three-dimensional tissue after the three-dimensional tissue scaffold is affixed to the support; and   measuring rotational displacement of at least a second portion of the three-dimensional tissue relative to the support over a period of time caused by functional activity of three-dimensional tissue or measuring strain of at least a second portion of the three-dimensional tissue relative to the support over a period of time caused by functional activity of the three-dimensional tissue.   
     
     
         2 .- 7 . (canceled) 
     
     
         8 . The method of  claim 1 , wherein measuring rotational displacement of at least the second portion of the three-dimensional tissue relative to the support over the period of time or measuring strain of at least the second portion of the three-dimensional tissue relative to the support over the period of time comprises obtaining images of at least the second portion of the three-dimensional tissue over the period of time. 
     
     
         9 .- 11 . (canceled) 
     
     
         12 . The method of  claim 1 , wherein the first portion of the three-dimensional tissue is disposed at a first end of the three-dimensional tissue and the at least the second portion of the three-dimensional tissue is disposed at a second end of the three-dimensional tissue opposite the first end of the three-dimensional tissue that is affixed to the support. 
     
     
         13 . The method of  claim 1 , further comprising:
 exposing the three-dimensional tissue scaffold to a solution containing fiducial markers such that the fiducial markers adhere to the three-dimensional tissue scaffold, or exposing the three-dimensional tissue to a solution containing fiducial markers that adhere to the three-dimensional tissue; and   wherein measuring rotational displacement of at least a second portion of the three-dimensional tissue relative to the support over a period of time or measuring strain of at least the second portion of the three-dimensional tissue relative to the support over the period of time comprises performing imaging of the fiducial markers in or on the at least a second portion of the three-dimensional tissue over the period of time.   
     
     
         14 .- 18 . (canceled) 
     
     
         19 . The method of  claim 1 , further comprising stimulating the three-dimensional tissue before, during, or before and during at least some of the measurement of the rotational displacement of at least the second portion of the three-dimensional tissue relative to the support over the period of time the measurement of the strain of at least the second portion of the three-dimensional tissue relative to the support over the period of time to cause a functional response in the three-dimensional tissue, to initiate the functional activity of the three-dimensional tissue, or to affect the functional activity of the three dimensional tissue. 
     
     
         20 .- 22 . (canceled) 
     
     
         23 . The method of  claim 1 , wherein the method is a method for identifying a compound that modulates tissue function; and
 wherein the method further comprises:
 contacting the three-dimensional tissue structure with a test compound, wherein measuring rotational displacement of at least a second portion of the three-dimensional tissue with respect to the support over a period of time or measuring strain of at least the second portion of the three-dimensional tissue relative to the support over the period of time includes:
 measuring a first rotational displacement of the at least the second portion of the three-dimensional tissue relative to the support in the presence of the test compound over a first period of time or measuring a first strain of at least the second portion of the three-dimensional tissue relative to the support in the presence of the test compound over a first period of time; and 
 measuring a second rotational displacement of the at least the second portion of the three-dimensional tissue relative to the support in the absence of the test compound over a second period of time or measuring a second strain of at least the second portion of the three-dimensional tissue relative to the support in the absence of the test compound over a second period of time; and 
 
 comparing the first rotational displacement of at least the second portion of the three-dimensional tissue or the first strain of at least the second portion of the three-dimensional tissue with the second rotational displacement of the at least the second portion of the three-dimensional tissue or the second strain of at least the second portion of the three-dimensional tissue, 
 wherein a modulation of the first rotational displacement or the first strain in the presence of the test compound as compared to the second rotational displacement or the second strain in the absence of the test compound indicates that the test compound modulates tissue function, thereby identifying a compound that modulates tissue function. 
   
     
     
         24 . The method of  claim 1 , wherein the method is a method for identifying a compound that is useful for treating or preventing a disease affecting tissue function, and
 wherein the method further comprises:
 contacting the three-dimensional tissue structure with a test compound, wherein measuring rotational displacement of the at least a second portion of the three-dimensional tissue with respect to the support over a period of time or measuring strain of at least the second portion of the three-dimensional tissue relative to the support over the period of time includes:
 measuring a first rotational displacement of the at least the second portion of the three-dimensional tissue relative to the support in the presence of the test compound over a first period of time or measuring a first strain of at least the second portion of the three-dimensional tissue relative to the support in the presence of the test compound over a first period of time; and 
 measuring a second rotational displacement of the at least the second portion of the three-dimensional tissue relative to the support in the absence of the test compound over a second period time or measuring a second strain of at least the second portion of the three-dimensional tissue relative to the support in the absence of the test compound over a second period of time; and 
 comparing the first rotational displacement of at least the second portion of the three-dimensional tissue or the first strain of at least the second portion of the three-dimensional tissue with the second rotational displacement of the at least the second portion of the three-dimensional tissue or the second strain of at least the second portion of the three-dimensional tissue, 
 
 wherein a modulation of the first rotational displacement or the first strain in the presence of the test compound as compared to the second rotational displacement or the second strain in the absence of the test compound indicates that the test compound modulates tissue function, thereby identifying a compound useful for treating or preventing a disease affecting tissue function. 
   
     
     
         25 . The method of  claim 1 , wherein the three-dimensional tissue is or includes a tissue engineered ventricle or a model of a ventricle, and wherein the method further comprises determining one or more ejection fractions for the three-dimensional tissue over time based on the measured rotational displacement of at least the second portion of the three-dimensional tissue relative to the support over the period of time, or based on the measured strain of at least the second portion of the three-dimensional tissue relative to the support over the period of time. 
     
     
         26 .- 33 . (canceled) 
     
     
         34 . A method comprising:
 providing or obtaining a three-dimensional tissue scaffold defining a lumen or a cavity, the tissue scaffold comprising one or more polymeric fibers each having a micron-scale or nanometer-scale diameter, at least some of the one or more polymeric fibers encircling the lumen or cavity at a helical angle with respect to a longitudinal axis of the lumen or cavity, at an azimuthal orientation with respect to the longitudinal axis of the lumen or cavity, or both;   providing or obtaining cells growing on or in the three-dimensional tissue scaffold to form a three-dimensional tissue defining the lumen or the cavity;   exposing the three-dimensional scaffold to a medium including fiducial markers;   obtaining images of an area including or adjacent to an opening of the lumen or cavity over a period of time;   performing particle imaging velocimetry on fiducial markers in the images of the area including or adjacent to the opening of the lumen or cavity; and   determining a volume, mass flux, or velocity of fluid flow out of or into the opening due to or affected by a functional activity of the three-dimensional tissue based on the particle imaging velocimetry.   
     
     
         35 . The method of  claim 34 , wherein the three-dimensional tissue is or includes a tissue engineered ventricle or a model of a ventricle, and wherein the method further comprises determining one or more ejection fractions for the three-dimensional tissue based, at least in part, on the determined volume, mass flux, or velocity of fluid flow out of or into the opening. 
     
     
         36 .- 39 . (canceled) 
     
     
         40 . The method of  claim 34 , wherein the fiducial markers are suspended in the medium. 
     
     
         41 .- 44 . (canceled) 
     
     
         45 . The method of  claim 34 , further comprising stimulating the three-dimensional tissue before, during, or before and during obtaining the images over the period of time, to initiate the functional activity of the three-dimensional tissue, or to affect the functional activity of the three-dimensional tissue. 
     
     
         46 . The method of  claim 34 , wherein the method is a method of evaluating the functional activity of the three-dimensional tissue in response to the stimulation. 
     
     
         47 . (canceled) 
     
     
         48 . (canceled) 
     
     
         49 . The method of  claim 34 , wherein the method is a method for identifying a compound that modulates tissue function;
 wherein the method further comprises contacting the three-dimensional tissue structure with a test compound;   wherein obtaining images of the area including or adjacent to the opening of the lumen or cavity over a period of time includes:
 obtaining first images of the area in the presence of the test compound over a first period of time; and 
 obtaining second images of the area in the absence of the test compound over a second period of time; 
   wherein determining a volume, mass flux, or velocity of fluid flow out of or into the opening due to or affected by a functional activity of the three-dimensional tissue based on the particle imaging velocimetry includes:
 determining a first volume, mass flux, or velocity of fluid flow out of or into the opening during the first period of time; and 
 determining a second volume, mass flux, or velocity of fluid flow out of or into the opening during the second period of time; and 
   wherein the method further comprises comparing the first volume, mass flux, or velocity of fluid flow with the second volume, mass flux, or velocity of fluid flow, wherein a modulation of the first volume, mass flux, or velocity of fluid flow in the presence of the test compound as compared to the second volume, mass flux, or velocity of fluid flow in the absence of the test compound indicates that the test compound modulates the tissue function.   
     
     
         50 . The method of  claim 34 , wherein the method is a method for identifying a compound that is useful for treating or preventing a disease affecting tissue function;
 wherein the method further comprises contacting the three-dimensional tissue structure with a test compound;   wherein obtaining images of the area including or adjacent to the opening of the lumen or cavity over a period of time includes:
 obtaining first images of the area in the presence of the test compound over a first period of time; and 
 obtaining second images of the area in the absence of the test compound over a second period of time; 
   wherein determining a volume, mass flux, or velocity of fluid flow out of or into the opening due to or affected by a functional activity of the three-dimensional tissue based on the particle imaging velocimetry comprises:
 determining a first volume, mass flux, or velocity of fluid flow out of or into the opening during the first period of time; and 
 determining a second volume, mass flux, or velocity of fluid flow out of or into the opening during the second period of time; and 
   wherein the method further comprises comparing the first volume, mass flux, or velocity of fluid flow and the second volume, mass flux, or velocity of fluid flow, wherein a modulation of the first volume, mass flux, or velocity of fluid flow as compared to the second volume, mass flux, or velocity of fluid flow indicates that the test compound modulates tissue function, thereby identifying a compound useful for treating or preventing a disease affecting tissue function.   
     
     
         51 .- 60 . (canceled) 
     
     
         61 . The method of  claim 1 , wherein the three-dimensional tissue structure comprises one or more of a heart, an artery, a blood vessel, a lymph node, a lymphatic vessel, an intestine, and an inner layer of a tongue. 
     
     
         62 . The method of  claim 1 , wherein the three-dimensional tissue scaffold includes:
 a first layer in which a first portion of the one or more polymeric fibers encircle the longitudinal axis of the lumen at a first helical angle with respect the longitudinal axis of the lumen or cavity; and   a second layer in which a second portion of the one or more polymeric fibers encircle the longitudinal axis of the lumen or cavity a second helical angle with respect to the longitudinal axis of the lumen or cavity, wherein a difference between the first helical angle and the second helical angle falls is less than 90°.   
     
     
         63 . (canceled) 
     
     
         64 . (canceled) 
     
     
         65 . The method of  claim 1 , wherein providing or obtaining the three-dimensional tissue scaffold comprises:
 rotating a reservoir holding a material comprising a polymer about a rotation axis to eject at least one jet of material from at least one orifice defined by an outer sidewall of the reservoir;   directing at least one flow of gas through a portion of the reservoir radially inward of the outer sidewall, the at least one flow of gas directed from an upstream first end of the reservoir to a downstream second end of the reservoir during rotation of the reservoir and ejection of the at least one jet of the material to form at least one micron or nanometer dimension polymeric fiber, the at least one flow of gas entraining the at least one micron or nanometer dimension polymeric fiber and forming a focused fiber deposition stream of the at least one micron or nanometer dimension polymeric fiber in a first direction, the first direction having an orientation of within 45 degrees of the rotation axis of the reservoir; and   collecting the focused fiber deposition stream on a target surface that is being rotated about a second rotation axis, the target surface having a shape corresponding to at least a portion of the three-dimensional tissue scaffold.   
     
     
         66 . (canceled)